Ultrasound-assisted electrochemical distinction of normal and cancerous cells

Abstract

A method for cancer diagnosis is disclosed. The method includes forming a plurality of cultured cells on an electrochemical biosensor placing the electrochemical biosensor in a medium solution comprising a cell culture solution of a plurality of biological cells, measuring a first electrochemical response from the electrochemical biosensor with the plurality of cultured cells, forming a plurality of stimulated cells on the electrochemical biosensor by ultrasonically stimulating of the plurality of cultured cells, measuring a second electrochemical response from the electrochemical biosensor with the plurality of stimulated cells, and detecting presence of cancer cells responsive to a difference between the first electrochemical response and the second electrochemical response being less than a threshold. Where, the first electrochemical response includes an electrochemical response of the plurality of cultured cells and the second electrochemical response includes an electrochemical response of the plurality of stimulated cells.

What is claimed is: 
     
       1. A method for detecting cancer cells, comprising:
 preparing an electrochemical-ultrasonic system, comprising:
 fabricating an electrochemical biosensor, the electrochemical biosensor comprising an integrated three-electrodes array patterned on a nanoroughened surface of a substrate, the integrated three-electrodes array comprising a working electrode, a counter electrode, and a reference electrode; 
 culturing a plurality of biological cells on the electrochemical biosensor comprising attaching the plurality of biological cells onto the working electrode by placing the electrochemical biosensor in a medium solution, the medium solution comprising a cell culture solution of the plurality of biological cells; 
 connecting the electrochemical biosensor with the plurality of cultured cells to an electrochemical stimulator-analyzer system, the electrochemical stimulator-analyzer system configured to measure electrochemical responses; and 
 exposing the electrochemical biosensor with the plurality of cultured cells to an ultrasonic system; 
 
 generating a plurality of microbubbles on the electrochemical biosensor with the plurality of cultured cells comprising electrolysis of the medium solution by applying an instantaneous electrical potential in the medium solution utilizing a cyclic voltammetry technique using the electrochemical stimulator-analyzer system, applying the instantaneous electrical potential in the medium solution comprising applying a DC signal with a voltage between −2 V and −0.5 V for a time duration less than 1 seconds on the biosensor with the plurality of cultured cells; 
 measuring a first electrochemical response from the electrochemical biosensor with the plurality of cultured cells using the electrochemical stimulator-analyzer system, the first electrochemical response comprising an electrochemical response of the plurality of cultured cells; 
 forming a plurality of stimulated cells on the electrochemical biosensor by applying ultrasonic waves to the plurality of cultured cells and the plurality of microbubbles using the ultrasonic system; 
 measuring a second electrochemical response from the electrochemical biosensor with the plurality of stimulated cells using the electrochemical stimulator-analyzer system, the second electrochemical response comprising an electrochemical response of the plurality of stimulated cells; and 
 detecting presence of cancer cells responsive to a difference between the first electrochemical response and the second electrochemical response being less than a threshold. 
 
     
     
       2. A method for detecting cancer cells, comprising:
 forming a plurality of cultured cells on an electrochemical biosensor by placing the electrochemical biosensor in a medium solution comprising a cell culture solution of a plurality of biological cells; 
 measuring a first electrochemical response from the electrochemical biosensor with the plurality of cultured cells, the first electrochemical response comprising a first cyclic voltammetry (CV) pattern comprising a first set of measured electrical currents versus a range of applied electrical potentials comprising a respective first current peak; 
 generating a plurality of microbubbles on the electrochemical biosensor with the plurality of cultured cells comprising electrolysis of the medium solution by applying an instantaneous electrical potential to the electrochemical biosensor with the plurality of cultured cells via a cyclic voltammetry technique using an electrochemical stimulator-analyzer system; 
 forming a plurality of stimulated cells on the electrochemical biosensor by applying ultrasonic waves to the plurality of cultured cells and the plurality of microbubbles using an ultrasonic system; 
 measuring a second electrochemical response from the electrochemical biosensor with the plurality of stimulated cells, the second electrochemical response comprising a second CV pattern comprising a second set of measured electrical currents versus the range of applied electrical potentials comprising a respective second current peak; and 
 detecting presence of cancer cells responsive to a difference between the first electrochemical response and the second electrochemical response being less than a threshold, the difference between the first electrochemical response and the second electrochemical response comprising a peak to peak 
 
       
         
           
             
               
                 
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       the peak to peak 
       
         
           
             
               
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       being defined by:
 peak to peak 
 
       
         
           
             
               
                 
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       where I USoff  comprises the first current peak and I USon  comprises the second current peak. 
     
     
       3. The method of  claim 2 , wherein detecting the presence of cancer cells comprises detecting presence of breast cancer cells responsive to the peak to peak 
       
         
           
             
               
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       being less than the threshold comprising a value of 0.15. 
     
     
       4. The method of  claim 3 , wherein:
 the threshold comprises a value of 0.05 for the ultrasonically stimulating of the plurality of cultured cells with an intensity of less than 1 W/cm 2  and a duration of less than 5 seconds, and 
 the threshold comprises a value of 0.15 for the ultrasonically stimulating of the plurality of cultured cells with an intensity of more than 1 W/cm 2  and a duration of less than 5 seconds. 
 
     
     
       5. The method of  claim 2 , wherein each of the measuring the first electrochemical response from the electrochemical biosensor with the plurality of cultured cells and measuring the second electrochemical response from the electrochemical biosensor with the plurality of stimulated cells comprises measuring an electrochemical response via the cyclic voltammetry technique using the electrochemical stimulator-analyzer system. 
     
     
       6. The method of  claim 5 , wherein the electrochemical stimulator-analyzer system comprises a potentiostat. 
     
     
       7. The method of  claim 2 , wherein forming the plurality of cultured cells on the electrochemical biosensor comprises:
 placing the electrochemical biosensor at the bottom of a chamber; and 
 filling the chamber with the medium solution. 
 
     
     
       8. The method of  claim 2 , wherein generating the plurality of microbubbles on the electrochemical biosensor with the plurality of cultured cells comprises applying a DC signal with a voltage between −2 V and −0.5 V for a time duration less than 1 seconds on the biosensor with the plurality of cultured cells using a potentiostat device. 
     
     
       9. The method of  claim 2 , wherein forming the plurality of stimulated cells on the electrochemical biosensor comprises:
 ultrasonically stimulating the plurality of cultured cells responsive to applying ultrasonic waves to the plurality of cultured cells on the electrochemical biosensor in the medium solution; and 
 inducing an acoustic cavitation in the plurality of cultured cells by the plurality of microbubbles responsive to applying ultrasonic waves to the plurality of microbubbles on the electrochemical biosensor in the medium solution. 
 
     
     
       10. The method of  claim 2 , wherein forming the plurality of stimulated cells on the electrochemical biosensor comprises:
 exposing the medium solution containing the electrochemical biosensor with the plurality of cultured cells to the ultrasonic system; and 
 applying ultrasonic waves to the medium solution with a frequency range between 10 KHz and 100 KHz for a time duration between 2s and 10 s to the plurality of cultured cells using the ultrasonic system. 
 
     
     
       11. The method of  claim 10 , wherein exposing the medium solution containing the electrochemical biosensor with the plurality of cultured cells to the ultrasonic system comprises placing an ultrasonic horn above the electrochemical biosensor with the plurality of cultured cells, the ultrasonic horn being connected to an ultrasonic generator. 
     
     
       12. The method of  claim 10 , wherein ultrasonic ultrasonically stimulating the plurality of cultured cells comprises applying ultrasonic waves using the ultrasonic system with an intensity between 0.5 w/cm 2  and 5 w/cm 2 .